Data Analysis Support
HOS activities using Nuclear Magnetic Resonance: NMR (Nuclear Magnetic Resonance) is used for characterization of drug formulation and used to study the behavior of drugs in various formulations, such as solution or solid state, and to understand how the formulation affects the stability and bioavailability of the drug. NMR spectroscopy also plays an important role in the development, quality control and characterization of biosimilars such as (i) Characterization of Higher Order Structure (HOS) including the 3D structure, dynamics and aggregation of proteins and other biologics. This information is critical for establishing the similarity between a biosimilar and its reference product, and for ensuring that the biosimilar has the same safety and efficacy profile as the reference product Further it is used for (ii) Comparison with the Reference Product: NMR can be used to compare the HOS of a biosimilar with that of its reference product. This comparison helps to establish the similarity between the two products, and to demonstrate that the biosimilar is highly similar to its reference product in terms of structure, function, and quality. (iii) Quality Control: NMR can be used to monitor the purity and quality of biosimilars during production and storage. This information is critical for ensuring that the biosimilars are free from contaminants and that they are stable over time (iv) Batch-to-Batch Consistency: NMR can be used to ensure batch-to-batch consistency of biosimilars. This is important for ensuring that each batch of a biosimilar has the same quality and performance as every other batch, and for ensuring that the biosimilar is consistently safe and effective for patients.
Analytical ultra centrifugation: Analytical ultracentrifugation (AUC) is a powerful analytical tool for studying the physical and chemical properties of biological molecules and biotherapeutics. For biotherapeutics AUC is used to study the higher order structure (HOS) of proteins, antibodies, and other biologics where some of the key applications of AUC are (i) Characterization of Macromolecular Assemblies: AUC can be used to study the size, shape, and stability of macromolecular assemblies, such as protein complexes, multimers, and aggregates. This information is important for understanding the interactions between proteins and other biologics, and for ensuring the stability of these assemblies over time (ii) Quality Control of Biotherapeutics: AUC can be used to monitor the quality of biotherapeutics during production and storage. This information is critical for ensuring that the biotherapeutics are free from contaminants and that they are stable over time (iii) Determination of Molecular Weight: AUC can be used to determine the molecular weight of proteins, antibodies, and other biologics. This information is important for characterizing the purity and quality of these biologics, and for understanding the physical properties of these molecules (iv) Characterization of Protein-Ligand Interactions: AUC can be used to study the binding of small molecules, such as drugs, to proteins and other biologics. This information is important for understanding the interactions between drugs and their target proteins, and for designing drugs that bind more specifically and with higher affinity to their target proteins.
Low resolution techniques: Low resolution techniques in biotherapeutics higher order structure characterization refer to analytical methods that provide information about the overall shape and size of a protein without resolving individual atoms or specific structural details. These techniques are particularly useful in characterizing the higher order structure (HOS) of biotherapeutics, such as monoclonal antibodies, recombinant proteins, and other biologics, where a comprehensive understanding of the protein's structural integrity is critical to ensuring its safety and efficacy. Some common low -resolution techniques used in HOS characterization include (i) Size exclusion chromatography (SEC): SEC separates proteins based on their size and shape, providing information about the molecular weight and oligomeric state of the protein. It is particularly useful in assessing the aggregation state of a protein, which is a critical quality attribute that can affect its stability and function (ii) Dynamic light scattering (DLS): DLS measures the size and distribution of particles in solution, including protein aggregates and complexes. It is a non-invasive technique that requires only small sample volumes, making it particularly useful in early-stage development and formulation studies. (iii) Differential scanning calorimetry (DSC): DSC measures the heat energy absorbed or released by a protein sample as it undergoes a temperature change. This technique provides information about the protein's thermal stability, including its melting temperature (Tm), which is a key indicator of its HOS integrity (iv) Circular dichroism (CD): CD measures the difference in the absorption of left- and right-circularly polarized light by a protein sample. It provides information about the protein's secondary structure, including alpha-helix, beta-sheet, and random coil content, and can be used to monitor conformational changes induced by changes in pH, temperature, or other environmental factors (v) Fourier transform infrared spectroscopy (FTIR) is a powerful analytical technique used in the characterization of biotherapeutics. It provides information about the higher order structure of proteins, including secondary structure, tertiary structure, and other structural characteristics. Low resolution FTIR is used to analyze the secondary structure of proteins, which includes the regular repeating patterns of the polypeptide backbone such as alpha-helices, beta-sheets, and random coils. FTIR measures the absorption of infrared radiation by protein molecules, which is sensitive to the different types of chemical bonds present in the protein backbone. The resulting spectrum can be deconvoluted using mathematical algorithms to estimate the relative contributions of different secondary structure components.Higher order structure analysis using FTIR involves the analysis of the protein's tertiary and quaternary structure, as well as other structural characteristics such as protein-protein interactions, hydrogen bonding, and solvent accessibility. This is typically accomplished using more advanced FTIR techniques such as attenuated total reflectance (ATR)-FTIR, two-dimensional (2D) FTIR, and synchrotron-based FTIR.